Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

Beat Zurbriggen; Michael J. Bailey; Merja E. Penttilä; Kaisa Poutanen; Matti Linko

doi:10.1016/0168-1656(90)90075-M

Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

Beat Zurbriggen, Michael J. Bailey, Merja E. Penttilä, Kaisa Poutanen, Matti Linko

Not published at Aalto University

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Abstract

Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars. © 1993.

Original language	English
Pages (from-to)	267-278
Number of pages	12
Journal	Journal of Biotechnology
Volume	13
Issue number	4
DOIs	https://doi.org/10.1016/0168-1656(90)90075-M
Publication status	Published - Mar 1990
MoE publication type	A1 Journal article-refereed

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@article{523e73a4cc8b447c99496bd38f6d7fd8,

title = "Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae",

abstract = "Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars. {\textcopyright} 1993.",

author = "Beat Zurbriggen and Bailey, {Michael J.} and Penttil{\"a}, {Merja E.} and Kaisa Poutanen and Matti Linko",

year = "1990",

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doi = "10.1016/0168-1656(90)90075-M",

language = "English",

volume = "13",

pages = "267--278",

journal = "Journal of Biotechnology",

issn = "0168-1656",

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TY - JOUR

T1 - Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

AU - Zurbriggen, Beat

AU - Bailey, Michael J.

AU - Penttilä, Merja E.

AU - Poutanen, Kaisa

AU - Linko, Matti

PY - 1990/3

Y1 - 1990/3

N2 - Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars. © 1993.

AB - Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars. © 1993.

U2 - 10.1016/0168-1656(90)90075-M

DO - 10.1016/0168-1656(90)90075-M

M3 - Article

SN - 0168-1656

VL - 13

SP - 267

EP - 278

JO - Journal of Biotechnology

JF - Journal of Biotechnology

IS - 4

ER -

Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

Abstract

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